Streptokinase (SK) is protein excreted by strains of β-hemolytic Streptococci. It is applied as treatment of life-threatening diseases thrombosed arteriovenous shunts, and thromboembolism. Streptokinase is naturally produced by several bacterial strains especially Streptococci. In connection with its broad application in the field of medicine, there is an urgent need to search for methods of its hyper production. Present study aimed at SK hyper production by mutagenesis of S. equisimilis strain. The wild bacterial strain S. equisimilis was subjected to random mutagenesis (UV irradiation and Ethidium bromide) for enhanced streptokinase production. The modified strain was assessed for the activity of streptokinase. The mutant S. equisimilis UV6 showed the maximum SK production and activity. By standardizing the effects of different factors like substrate (CSL), carbon source, salt concentration, temperature and pH an optimum fermentation protocol for SK production was established. The higher SK production was observed in mutant strain UV6 601 U mL-1 with increased activity up to 400 %. The SK was purified and its kinetics data indicated improved activity as its Km value was low as compared to wild SK. The SK gene from mutant UV6 was amplified and cloned in E. coli using pTZ57R/T vector. The gene was sequenced and compared with the wild SK gene to check the genetic difference between wild and mutant strain. In our study, the comparative sequence analysis revealed 26 point mutations, among these 3 were transitions (A to G) while the remaining 23 were transversions (A/G to C/T) in coding region of sk gene. Moreover, 3 amino acid deletions at the C terminal region of SK protein were predicted due to non-sense mutation in coding region (resulting in a stop codon after residue 437). The activity of any protein is direcly linked with amino acid composision. in our studies, the in-silico analyses predicted that amino acid residues at position 22, 62, 64 and 80 could be accountable in SK hyperactivity.
جے کوئی تینوں آوے چج عشقے توں جا بیبا بھج جھوٹا ایں تے چپ کر بہہ سچا ہیں بن بدل گج جے اوہ جاوے نظریں آ ویکھ کے اکھیاں جاون رج عظمت کالی کملی دی جو رکھدی اے عیباں دی لج ڈِھڈ دی گل سناویں ہا اینویں لا نہ جھوٹے پج جنہوں عشق حضورؐ دا اے اونہاں دے نت ہوندے حج
This research is based on the correction of a common mistake that is used in many of the books and scientific field. It has been spread among many of researchers these days that the Prophet Mohammad’s (SAW) Companions stripped the Holy Qur’an off the dots and diacritics to cover the aspects of the Holy Qur’anic readings. In the introduction, the researcher talks about the importance of the topic and the reason why it has been chosen. In the first chapter covers the meaning of the terms that the research is based on such as: dots, inserting dots, diacritics and stripping. The second chapter, the researcher mentions the implications of stripping and clarifies its meaning and its effect on the understanding of the researchers at the present time. In the third chapter, the researcher talks about the history of dots, where he mentions and analyzes a number of old inscriptions that are considered as the basis from which Arabic language was developed. In conclusion, the researcher refers to the most important findings and recommendations.
This PhD thesis presents details of electrode modification and its application as a sensitive electrochemical platform for efficient water splitting and traces level detection of drugs. For water splitting the reaction for oxygen evolution has unfavourable thermodynamics since it needs energy to proceed. Lately scientists are focused to develop cheap oxygen evolving catalysts that are abundantly present in nature and have succeeded as well. But stability and efficiency of these catalysts still demands improvements. In this respect a part of the current work is focused on the development of an industrially favourable material that is durable and suitable for oxygen evolution reaction (OER) at low values of overpotential. Herein the synthesis of a novel and stable alkyl amine ligands containing NiFe complex material is reported, which displays remarkably high OER performance and extraordinary stability for OER stretching over days at varying temperature conditions under alkaline conditions. The powdered form of catalyst was found to give signal of OER at overpotential of 290 mV for consecutive 14 hours. The presence of binder in the preparation method of the powdered catalyst caused to affect its conductivity. Therefore, the catalyst was directly grown on nickel foam electrode which demonstrated improved water splitting performance as evidenced by 230 mV overpotential and stability upto consecutive 20 hours. After modification, the electrode was developed into a sensor for drugs analysis. Three different nanobiosensors were prepared using graphene oxide, multiwalled carbon nanotubes and gold and silver nanoparticles using screen printed electrode. Nanomaterials are associated with favorable characteristics including large surface area and rare electrical properties. So, nanomaterial-enriched deoxyribonucleic acid DNA sensors were developed for sensing the DNA binding events of a number of anticancer drugs. The results revealed decrease in signals response of DNA bases (adenine and guanine) due to the damage of the oxidizable moieties of these electroactive bases. The sensors demonstrated good values of sensitivity, selectivity, reproducibility, fast detection ability and to analyse drugs in its pharmaceutical dosage.